Assessment of liposome disruption to quantify drug delivery in vitro

Efficient liposome disruption inside the cells is a key for success with any type of drug delivery system. The efficacy of drug delivery is currently evaluated by direct visualization of labeled liposomes internalized by cells, not addressing objectively the release and distribution of the drug. Here, we propose a novel method to easily assess liposome disruption and drug release into the cytoplasm. We propose the encapsulation of the cationic dye Hoechst 34,580 to detect an increase in blue fluorescence due to its specific binding to negatively charged DNA. For that, the dye needs to be released inside the cell and translocated to the nucleus. The present approach correlates the intensity of detected fluorescent dye with liposome disruption and consequently assesses drug delivery within the cells.

Como é que os casais experienciam o relacionamento íntimo após o diagnóstico de cancro?

Dissertação de mestrado integrado em Psicologia

Empatia no desenvolvimento típico: relação entre autorrelatos e comportamentos empáticos

Dissertação de mestrado integrado em Psicologia

Resolução da ambivalência em terapia narrativa

Dissertação de mestrado integrado em Psicologia

Influência da corrosão no comportamento ao arrancamento de fibras de aço no betão auto-compactável

Quando adicionadas ao betão, a possibilidade de corrosão das fibras de aço , devido à penetração de cloretos e ação da carbonatação, é um parâmetro de durabilidade no betão auto-compactável reforçado com fibras de aço (BACRFA) que requer especial atenção. No BACRFA fendilhado, a corrosão tende a ocorrer nas fibras de aço que atravessam as fissuras, o que pode afetar o seu comportamento à flexão. Deste modo, o estudo do comportamento interfacial fibra/matriz é importante para a compreensão dos efeitos da corrosão das fibras de aço no comportamento mecânico e na durabilidade das estruturas de BACRFA. Com o intuito de avaliar estes efeitos, no presente trabalho desenvolveu-se um ensaio experimental de arrancamento de fibras de aço em BAC fendilhado, sujeitas previamente à ação da corrosão por exposição a um ambiente agressivo de cloretos, sendo a corrosão avaliada por intermédio de técnicas eletroquímicas. Os ensaios eletroquímicos realizados revelaram que a probabilidade de corrosão das fibras de aço num ambiente com cloretos é superior a 90%. No mesmo ambiente, a susceptibilidade para a corros ão das fibras de aço que atravessam uma pseudo-fissura, aumenta com a abertura de fissura provocando um aumento da resistência ao arranque da fibra em BAC fissurado

Modelos de apoio às decisão na conceção e planeamento de um armazém

Programa Doutoral em Engenharia Industrial e de Sistemas.

Intervention program on adolescent’s creativity representations and academic motivation

Creativity and its promotion are widespread concerns in education. However, few efforts have been made to implement intervention programs designed to promote creativity and other related aspects (e.g., academic motivation). The Future Problem Solving Program International (FPSPI), aimed for training creativity representations and creative problem solving skills in young people, has been one of the most implemented programs. This intervention’s materials and activities were adapted for Portuguese students, and a longitudinal study was conducted. The program was implemented during four months, in weekly sessions, by thirteen teachers. Teachers received previous training for the program and during the program’s implementation. Intervention participants included 77 Basic and Secondary Education students, and control participants included 78 equivalent students. Pretest-posttest measures of academic motivation and creativity representations were collected. Results suggest a significant increase, in the intervention group, in motivation and the appropriate representations of creativity. Practical implications and future research perspectives are presented.

Data quality in biofilm high-throughput routine analysis: intralaboratory protocol adaptation and experiment reproducibility

Biofilm research is growing more diverse and dependent on high-throughput technologies and the large-scale production of results aggravates data substantiation. In particular, it is often the case that experimental protocols are adapted to meet the needs of a particular laboratory and no statistical validation of the modified method is provided. This paper discusses the impact of intra-laboratory adaptation and non-rigorous documentation of experimental protocols on biofilm data interchange and validation. The case study is a non-standard, but widely used, workflow for Pseudomonas aeruginosa biofilm development, considering three analysis assays: the crystal violet (CV) assay for biomass quantification, the XTT assay for respiratory activity assessment, and the colony forming units (CFU) assay for determination of cell viability. The ruggedness of the protocol was assessed by introducing small changes in the biofilm growth conditions, which simulate minor protocol adaptations and non-rigorous protocol documentation. Results show that even minor variations in the biofilm growth conditions may affect the results considerably, and that the biofilm analysis assays lack repeatability. Intra-laboratory validation of non-standard protocols is found critical to ensure data quality and enable the comparison of results within and among laboratories.

A redefinição das condições estruturais da infância e a crise económica em Portugal

Nos últimos 6 anos (desde 2008), a Europa tem estado mergulhada numa crise económica e financeira que atingiu, em particular, as economias mais vulneráveis, nomeadamente as dos países do Sul. Enfrentando crises de dívida soberana, a maioria dos países do Sul da Europa – Portugal, Espanha, Grécia, Chipre, Itália e Irlanda – recorreram à intervenção da União Europeia e do FMI. A austeridade tem sido a receita seguida em todos esses países, com constrangimentos no uso de dinheiro público, restrições nas políticas sociais, despedimentos na administração pública, redução dos salários nominais e aumento de impostos sobre a população ativa. Ao mesmo tempo, reformas estruturais têm sido anunciadas pelos diferentes governos, com o objetivo proclamado de diminuir a dívida pública e de aumentar o crescimento económico. Porém, estes planos são, por norma, acompanhados de medidas que reduzem a capacidade financeiras das famílias, designadamente, além dos cortes salariais, as reduções e restrições nos principais benefícios sociais e políticas familiares específicas (como, por exemplo, os abonos de família). As consequências que esta “economia global” tem tido nas vidas das crianças portuguesas são significativas e acreditamos ser fundamental refletir sobre os modos atuais de estruturação da infância a partir desta realidade e das implicações concretas que têm tido nas condições de vida das crianças. As condições estruturais em que a categoria geracional da infância é construída, em cada momento histórico, são geradoras de possibilidades reais de vida das crianças. As relações entre gerações não são sempre uniformes; pelo contrário, modificam-se continuamente. Diferentes condições de estruturação (isto é, em termos giddensianos, diferentes modos de regulação e de alocação de recursos) da vida económica, social e cultural geram distintos processos de “generatividade” (Honnig, 2009), isto é, modos de relação e mutua configuração entre gerações, especialmente entre adultos e crianças. A crise económica alterou as relações entre gerações. Um indicador evidente disto mesmo é a forte redução do número de crianças, através da descida do índice sintético de fertilidade nos anos da crise (muito embora esse índice tenha uma tendência de descida anterior): 1,37 em 2008; 1,28 em 2012 (INE, 2014). Mas também nos domínios da administração simbólica da infância, a crise económica produz os seus efeitos: a redução dos direitos, sendo generalizada, afeta especial e especificamente a infância. O nosso ponto é o de que a infância sofre um processo de redefinição, através da restrição às condições de cidadania da infância, por efeito da crise e da forma de gestão política que ela assumiu na europa, especialmente em Portugal. As reflexões sobre este efeito de redefinição da infância serão feitas a partir de duas dimensões centrais: uma, de análise de dados gerais sobre as políticas atuais que têm vindo a diminuir direitos sociais, a aumentar os níveis de pobreza e a ameaçar direitos fundamentais como educação e saúde, visibilizando os seus impactos nas vidas das crianças. Uma segunda dimensão discutirá dados recolhidos com crianças de modo a tentar caracterizar, a partir dos seus pontos de vista, os significados e impactos da crise nas suas vidas.

Personalidade e bem-estar subjetivo na excelência profissional: validação de escalas junto de gestores portugueses

O presente estudo teve como objetivo a validação da Satisfaction With Life Scale (SWLS; Diener, Emmons, Larsen, & Griffin, 1985) e da Core Self-Evaluations Scale (CSES; Judge, Erez, Bono, & Thoresen, 2003) junto de gestores empresariais portugueses (N = 108). A escala SWLS avalia a componente cognitiva do bem-estar psicológico, isto é, a satisfação percebida face à qualidade das condições de vida. A CSES, centrada na valoração básica que a pessoa faz do seu self, abarca quatro dimensões de personalidade: autoestima, autoeficácia generalizada, neuroticismo e lócus de controlo. Os resultados encontrados sugerem que ambos os instrumentos apresentam índices adequados de precisão e que os constructos iniciais definidos pelos autores se mantêm parcialmente na estrutura dimensional das escalas nesta amostra portuguesa, havendo maior proximidade na SWLS. Foi observada, ainda, uma correlação positiva e estatisticamente significativa entre traços de personalidade avaliados pela CSES e a satisfação com a vida avaliada pela SWLS, em particular relativamente aos itens reportados à perceção de autoeficácia.

Reconfiguração e novas funcionalidades para um servidor REST ONVIF: serviço de IO e configuração de descoberta

Dissertação de mestrado em Engenharia de Telecomunicações e Informática

Exploring female mice interstrain differences relevant for models of depression

Depression is an extremely heterogeneous disorder. Diverse molecular mechanisms have been suggested to underlie its etiology. To understand the molecular mechanisms responsible for this complex disorder, researchers have been using animal models extensively, namely mice from various genetic backgrounds and harboring distinct genetic modifications. The use of numerous mouse models has contributed to enrich our knowledge on depression. However, accumulating data also revealed that the intrinsic characteristics of each mouse strain might influence the experimental outcomes, which may justify some conflicting evidence reported in the literature. To further understand the impact of the genetic background, we performed a multimodal comparative study encompassing the most relevant parameters commonly addressed in depression, in three of the most widely used mouse strains: Balb/c, C57BL/6, and CD-1. Moreover, female mice were selected for this study taken into account the higher prevalence of depression in women and the fewer animal studies using this gender. Our results show that Balb/c mice have a more pronounced anxious-like behavior than CD-1 and C57BL/6 mice, whereas C57BL/6 animals present the strongest depressive-like trait. Furthermore, C57BL/6 mice display the highest rate of proliferating cells and brain-derived neurotrophic factor (Bdnf) expression levels in the hippocampus, while hippocampal dentate granular neurons of Balb/c mice show smaller dendritic lengths and fewer ramifications. Of notice, the expression levels of inducible nitric oxide synthase (iNos) predict 39.5% of the depressive-like behavior index, which suggests a key role of hippocampal iNOS in depression. Overall, this study reveals important interstrain differences in several behavioral dimensions and molecular and cellular parameters that should be considered when preparing and analyzing experiments addressing depression using mouse models. It further contributes to the literature by revealing the predictive value of hippocampal iNos expression levels in depressive-like behavior, irrespectively of the mouse strain.

A breakthrough on Amanita phalloides poisoning: an effective antidotal effect by polymyxin B

Amanita phalloides is responsible for more than 90 % of mushroom-related fatalities, and no effective antidote is available. a-Amanitin, the main toxin of A. phalloides, inhibits RNA polymerase II (RNAP II), causing hepatic and kidney failure. In silico studies included docking and molecular dynamics simulation coupled to molecular mechanics with generalized Born and surface area method energy decomposition on RNAP II. They were performed with a clinical drug that shares chemical similarities to a-amanitin, polymyxin B. The results show that polymyxin B potentially binds to RNAP II in the same interface of a-amanitin, preventing the toxin from binding to RNAP II. In vivo, the inhibition of the mRNA transcripts elicited by a-amanitin was efficiently reverted by polymyxin B in the kidneys. Moreover, polymyxin B significantly decreased the hepatic and renal a-amanitin-induced injury as seen by the histology and hepatic aminotransferases plasma data. In the survival assay, all animals exposed to a-amanitin died within 5 days, whereas 50 % survived up to 30 days when polymyxin B was administered 4, 8, and 12 h post-a-amanitin. Moreover, a single dose of polymyxin B administered concomitantly with a-amanitin was able to guarantee 100 % survival. Polymyxin B protects RNAP II from inactivation leading to an effective prevention of organ damage and increasing survival in a-amanitin-treated animals. The present use of clinically relevant concentrations of an already human-use-approved drug prompts the use of polymyxin B as an antidote for A. phalloides poisoning in humans.

Autoimmune diseases and pregnancy: analysis of a series of cases

BACKGROUND: An autoimmune disease is characterized by tissue damage, caused by self-reactivity of different effector mechanisms of the immune system, namely antibodies and T cells. All autoimmune diseases, to some extent, have implications for fertility and obstetrics. Currently, due to available treatments and specialised care for pregnant women with autoimmune disease, the prognosis for both mother and child has improved significantly. However these pregnancies are always high risk. The purpose of this study is to analyse the fertility/pregnancy process of women with systemic and organ-specific autoimmune diseases and assess pathological and treatment implications. METHODS: The authors performed an analysis of the clinical records and relevant obstetric history of five patients representing five distinct autoimmune pathological scenarios, selected from Autoimmune Disease Consultation at the Hospital of Braga, and reviewed the literature. RESULTS: The five clinical cases are the following: Case 1-28 years old with systemic lupus erythematosus, and clinical remission of the disease, under medication with hydroxychloroquine, prednisolone and acetylsalicylic acid, with incomplete miscarriage at 7 weeks of gestation without signs of thrombosis. Case 2-44 years old with history of two late miscarriages, a single preterm delivery (33 weeks) and multiple thrombotic events over the years, was diagnosed with antiphospholipid syndrome after acute myocardial infarction. Case 3-31 years old with polymyositis, treated with azathioprine for 3 years with complete remission of the disease, took the informed decision to get pregnant after medical consultation and full weaning from azathioprine, and gave birth to a healthy term new-born. Case 4-38 years old pregnant woman developed Behcet's syndrome during the final 15 weeks of gestation and with disease exacerbation after delivery. Case 5-36 years old with autoimmune thyroiditis diagnosed during her first pregnancy, with difficult control over the thyroid function over the years and first trimester miscarriage, suffered a second miscarriage despite clinical stability and antibody regression. CONCLUSIONS: As described in literature, the authors found a strong association between autoimmune disease and obstetric complications, especially with systemic lupus erythematosus, antiphospholipid syndrome and autoimmune thyroiditis.

An efficient chronic unpredictable stress protocol to induce stress-related responses in C57BL/6 mice

Exposure to chronic stress can have broad effects on health ranging from increased predisposition for neuropsychiatric disorders to deregulation of immune responses. The chronic unpredictable stress (CUS) protocol has been widely used to study the impact of stress exposure in several animal models and consists in the random, intermittent, and unpredictable exposure to a variety of stressors during several weeks. CUS has consistently been shown to induce behavioral and immunological alterations typical of the chronic stress-response. Unfortunately C57BL/6 mice, one of the most widely used mouse strains, due to the great variety of genetically modified lines, seem to be resistant to the commonly used 4-week-long CUS protocol. The definition of an alternative CUS protocol allowing the use of C57BL/6 mice in chronic stress experiments is a need. Here, we show that by extending the CUS protocol to 8?weeks is possible to induce a chronic stress-response in C57BL/6 mice, as revealed by abrogated body weight gain, increased adrenals weight, and an overactive hypothalamic-pituitary-adrenal axis with increased levels of serum corticosterone. Moreover, we also observed stress-associated behavioral alterations, including the potentiation of anxious-like and depressive-like behaviors and a reduction of exploratory behavior, as well as subtle stress-related changes in the cell population of the thymus and of the spleen. The present protocol for C57BL/6 mice consistently triggers the spectrum of CUS-induced changes observed in rats and, thus, will be highly useful to researchers that need to use this particular mouse strain as an animal model of neuropsychiatric disorders and/or immune deregulation related to CUS.